In general, any new information about the culture is integrated by adding new balance-like equations (e.g. Equation (REF )) or changing the bounds of the reaction fluxes (e.g. Equation (REF )) [1]}.
For example, we can define a new constraint that accounts for physical and spatial restrictions resulting from the limited resources accessible to the cell (\(\emph {e.g.}\)  cell volume, membrane area, enzyme solubility, proteome, etc.) [2]}, [3]}, [4]}, [5]}, [6]}.
It can be formulated as:
\(\sum _j^N (a_j^+r_j^++ a_j^-r_j^-) \le 1\) 
